人胸腺素α1基因在毕赤酵母中的融合表达  

Construction and Expression of Recombinant Human Thymosin α1 Fusion Gene in Pichia pastoris

作　　者：黄雯[1] 李兆育[1] 金礼吉[1] 安利佳[1] 

机构地区：[1]大连理工大学生物工程系,大连116012

出　　处：《遗传》2002年第6期679-683,共5页Hereditas（Beijing)

基　　金：辽宁省科学技术基金(No.99305001)资助

摘　　要：根据已知人胸腺素1(humanThymosinα1,hTα1)的氨基酸序列和毕赤酵母偏爱密码子,人工合成了hTα1基因。用合成的hTα1基因取代核糖体蛋白(ribosomalprotein,RP)基因5′端的84个碱基,组成重组基因hTα1-RP,连接到质粒pPIC3.5K上,构建表达质粒pPIC3.5K/hTα1-RP。表达质粒用电激法转化到毕赤酵母GS115菌株中。甲醇诱导表达融合蛋白,SDS-PAGE和Western印迹结果证明,重组基因hTα1-RP在酵母中得到了表达,表达量约为25mg/L。The human Thymosin α1 (hTα1) gene was synthesized according to the optimal codons of Pichia pastoris and was fused in 5' terminal of ribosomal protein (RP) gene using over lapping polymerase chain reaction.The fusion gene was inserted into expression vector of pPIC3.5K and was transformed into HIS4 mutant strain GS115 by electroporation.Both SDS-PAGE and Western blot indicated that this fusion protein was expressed.The expression level was about 25mg/L.

关 键 词：人胸腺素α1 基因 毕赤酵母 融合表达 融合蛋白 

分 类 号：TQ464[化学工程—制药化工]